The Mechanism Research Of Scoparone Inhibits Inflammatory Response And Alleviates Hepatic Fibrosis By Regulating TLR-4/NF-?B Signal Pathway

Objects:To investigate the anti-hepatic fibrosis effect of Scoparone?SCO?in vitro and in vivo,and the role of SCO on TLR-4/NF-?B signaling pathway in inhibiting progress of hepatic fibrosis was discussed.Aim to find a effective targets and ideas for the treatment of liver fibrosis.Methods:CCl₄ and transforming growth factor-?1?TGF-?1?were used to successfully establish hepatic fibrotic models in rat and hepatic stellate cell-T6?HSC-T6?.In vivo experiments,60 male SD rats were selected and divided into 6 groups,named normal group,model group,colchicine group?0.12mg/kg?,SCO high,medium and low dose groups?200,100,50 mg/kg?.HE staining was observed to evaluate the damage degree of liver fibrosis,The enzymes activity level of ALT,AST,ALP,?-GT,SOD,GSH-Px in rat serum were detected by biochemical method;TBIL and MDA in serum were detected by microplate method;Serum collagen deposition factor of HA?LN?PC-III and IV-C were detected by Enzyme-linked immune sorbent assay?ELISA?;ELISA assays were used to calculate the contents of TNF-??IL-6 and IL-1?in serum;The expression of TLR-4/NF-?B pathway related protein were dected by Western Blot and Q-PCR.In vitro experiments,HSC-T6 cells were divided into Control group,Model group?10 ng/m L?and SCO high,medium and low dose groups?80,60,40?M?,Immunofluorescence experiments were used to observe the expression of?-SMA and NF-?B protein after HSC-T6 was induced by TGF-?1;Woung-healing were used to detect cell migration;The levels of TLR-4/NF-?B pathway in HSC-T6 cells were dected by using Western Blot and Q-PCR.Results:?1?SCO had a protective effect on hepatic fibrosis rats induced by CCl₄.The protective effect was best,when the concentration was 200 mg/kg.?2?SCO can reduce the expression of TLR-4,My D88,p-NF-?B protein on TLR-4/NF-?B pathway in liver fibrosis rats,and significantly reduce the downstream factors TNF-?,IL-6,IL-1?contents.?3?SCO inhibited the proliferation of HSC-T6 cells activated by TGF-?1,and its IC50 is 60.12?M.?4?SCO can reduce the expression of TLR-4,My D88,p-NF-?B protein on TLR-4/NF-?B pathway HSC-T6 cells,and significantly decrease the downstream factors TNF-?,IL-6,IL-1?contents.?5?After SCO inhibited the TLR-4/NF-?B pathway in vitro and in vivo,the expression of?-SMA and TGF-?1 in liver fibrosis-related proteins were significantly reduced.Conclusions:SCO can block the TLR-4/NF-?B pathway in vitro and in vivo to exert anti-fibrotic effects.